CN111607622B - Process method for producing 3-hydroxy butanone by using wheat B starch - Google Patents
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Abstract
The invention provides a process method for producing 3-hydroxy butanone by using wheat B starch, belonging to the technical field of deep processing of agricultural products. The specific process method comprises the following steps: preparing sugar from wheat B starch to obtain starch sugar liquid, preparing a fermentation culture medium by using the starch sugar liquid as a raw material, and inoculating a 3-hydroxy butanone production strain into the fermentation culture medium for fermentation to produce the 3-hydroxy butanone. By adopting the process method, the production cost of the 3-hydroxy butanone can be effectively reduced, and the additional value of the wheat B starch is effectively increased, so that the process method has good practical application value.
Description
Technical Field
The invention belongs to the technical field of deep processing of agricultural products, and particularly relates to a process method for producing 3-hydroxy butanone by using wheat B starch.
Background
The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
The 3-hydroxy butanone (acetoin), also known as acetoin and acetyl methyl methanol, is an edible spice and an important C4 platform compound which are widely applied and can be widely applied to the fields of food, medicine, chemical industry, tobacco and the like. At present, 3-hydroxy butanone products on the market are mainly produced by a chemical synthesis method, and the price is about 10-15 ten thousand yuan/ton. The chemical method has limited raw material sources and more byproducts, and the product does not conform to the consumption concept of nature, green and safety, thereby limiting the application and development of the product in the food industry. In addition, the purity of the 3-hydroxy butanone produced by a chemical method is low, so that the application and popularization of the 3-hydroxy butanone as a platform compound in the industries of medicine, chemical engineering and the like are limited. The 3-hydroxy butanone is a metabolite of various microorganisms, and can be produced by microbial transformation with glucose as a raw material. The 3-hydroxy butanone produced by biotransformation has the advantages of abundant raw material sources, high product purity, high product safety and the like, the market price of the product is 2 times (25-30 ten thousand yuan/ton) of that of a chemical method product, and the added value of the product is higher. The inventor carries out biotechnology to produce 3-hydroxy butanone for years, and the screened bacillus subtilis 3-hydroxy butanone has high yield and better industrial application potential (Pingxiang and the like, a bacillus subtilis producing high-purity 3-hydroxy butanone, Chinese patent ZL2007100134025, CN101016530B, 2007; Pingxiang and the like, a bacillus producing 3-hydroxy butanone and application thereof, Chinese patent ZL201310289934.7, 2013; Van Yixiao and the like, screening and product analysis of a strain producing 3-hydroxy butanone, and food fermentation industry 2012,38(11): 42-46.). The technology for producing 3-hydroxy butanone by biotransformation reported at present basically takes glucose as raw material, in addition, organic nitrogen sources such as yeast extract and corn steep liquor, inorganic nitrogen sources such as ammonium sulfate and urea, and a small amount of other inorganic salts are also required to be added into a fermentation culture medium, and the raw material cost accounts for about 60-70% of the total production cost.
Wheat gluten production is one of the major deep processing modes of wheat, wherein the protein content of the wheat is about 12-13%, and the starch content is 65-70%, so that a large amount of wheat starch is co-produced in the process of producing wheat gluten. The wheat starch is divided into starch A and starch B according to the difference of particle sizes, wherein the starch A has large particles, is relatively easy to separate by centrifugation, and can be further refined to be sold as commercial starch. The starch B is starch with small particles, and during production, non-starch components such as pentosan, residual protein and the like in the wheat starch slurry form a three-phase centrifuge separation intermediate phase component together, and the starch component in dry substances of the component accounts for about 80 percent, has high viscosity and strong water holding capacity, and is difficult to increment and dispose. The inventor finds that the B starch is mainly used for alcoholic fermentation in the current production or sold as feed after being dried, and the added value of the product is low.
Disclosure of Invention
Aiming at the problem of low-value utilization of wheat B starch generated in the existing wheat gluten production process, the invention provides a process method for producing 3-hydroxy butanone by utilizing the wheat B starch. Specifically, the wheat B starch produced in the production process of the wheat gluten is used for fermentation production of the 3-hydroxy butanone after being used for preparing sugar. By adopting the process method, the production cost of the 3-hydroxy butanone can be effectively reduced, and the additional value of the wheat B starch is effectively increased, so that the process method has good practical application value.
In order to achieve the technical purpose, the technical scheme of the invention is as follows:
in a first aspect of the invention, there is provided the use of wheat B starch in the fermentative production of 3-hydroxybutanone.
In a second aspect of the present invention, there is provided a process for producing 3-hydroxybutanone from wheat B starch, the process comprising: preparing sugar from wheat B starch to obtain starch sugar liquid, preparing a fermentation culture medium by using the starch sugar liquid as a raw material, and inoculating a 3-hydroxy butanone production strain into the fermentation culture medium for fermentation to produce the 3-hydroxy butanone.
Furthermore, the method for preparing sugar from wheat B starch is not limited, such as a common sugar preparation process by a two-enzyme method;
and further, adding water into the wheat B starch for size mixing to obtain starch slurry, controlling the concentration of the starch slurry to be 10-15 Be, and preparing a starch sugar solution by adopting a double-enzyme method sugar preparation process.
Further, in the fermentation medium, the initial concentration of glucose is controlled to be 120-160 g/L;
further, the fermentation medium also comprises an inorganic nitrogen source; still further, the inorganic nitrogen source may be urea. The research of the invention finds that yeast extract is not needed to be added when the starch syrup B is used as the raw material for 3-hydroxy butanone fermentation, and the yield and the conversion rate of the 3-hydroxy butanone are obviously higher than those of glucose.
In a third aspect of the invention, the 3-hydroxy butanone prepared by the process method for producing the 3-hydroxy butanone by using the wheat B starch is provided.
The beneficial technical effects of one or more technical schemes are as follows:
1) the part B of starch separated in the production process of wheat gluten powder accounts for about 40 percent of dry matter, about 80 percent of starch in the dry matter, and other parts also contain protein, fat, crude fiber, ash and the like. Researches show that when the 3-hydroxy butanone is used as a raw material for fermentation, only a small amount of inorganic nitrogen source is needed to be added, and expensive organic nitrogen sources such as yeast extract and the like do not need to be added. Meanwhile, the cost of the starch B is far lower than that of glucose, so that the raw material cost for producing the 3-hydroxy butanone can be greatly reduced by utilizing the starch B of the wheat;
2) 3-hydroxy butanone and water azeotropy, can collect 3-hydroxy butanone in the fermented solution through reduced pressure distillation or multiple-effect concentration, the B starch raw material brings other unfermented ingredients such as pentosan and the like to be enriched in the bottom distillate, and compared with glucose raw material, the extraction cost of 3-hydroxy butanone is not increased. While the bottom fraction can be further disposed of as feed.
3) The starch B obtained by separating the wheat starch is used for producing the 3-hydroxy butanone, the added value of the product is high, and the high added value utilization of the wheat starch B is realized, so that the wheat starch B has good value of practical application.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise. It is to be understood that the scope of the invention is not to be limited to the specific embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention.
In one embodiment of the invention, the use of wheat B starch in the fermentative production of 3-hydroxybutanone is provided. The invention reports that the starch sugar solution of wheat B starch is used as a culture medium component for producing 3-hydroxy butanone by fermentation of a production strain for the first time, tests prove that the fermentation production of 3-hydroxy butanone can be realized only by adding a small amount of inorganic nitrogen source into the culture medium, and the yield and the conversion rate of the 3-hydroxy butanone are high.
In another embodiment of the present invention, there is provided a process for producing 3-hydroxybutanone from wheat B starch, comprising: preparing sugar from wheat B starch to obtain starch sugar liquid, preparing a fermentation culture medium by using the starch sugar liquid as a raw material, and inoculating a 3-hydroxy butanone production strain into the fermentation culture medium for fermentation to produce the 3-hydroxy butanone.
In yet another embodiment of the present invention, the wheat B starch may be selected from B starch slurry separated during vital gluten processing; or selecting the B starch product prepared by directly drying the B starch;
in another embodiment of the present invention, the wheat B starch may be:
centrifuging fresh wheat starch slurry generated in the production process of wheat gluten by using a centrifuge at the rotating speed of 3000rpm for 2 seconds, removing supernatant liquid, and collecting the middle flowable slurry part;
or separating wheat gluten from the intermediate phase separated in the wheat gluten production process of the three-phase horizontal decanter centrifuge to obtain B starch slurry;
or commercial starch B (dry powder) from wheat starch manufacturers.
In another embodiment of the invention, the method for preparing sugar from wheat B starch is not limited, such as a commonly used "two-enzyme method" sugar preparation process;
in another specific embodiment of the invention, the wheat B starch is added with water and is subjected to size mixing to obtain starch slurry, the concentration of the starch slurry is controlled to be 10-15 Be, and then a starch sugar solution is prepared by adopting a double-enzyme method sugar preparation process.
In yet another embodiment of the present invention, the fermentation medium further comprises an inorganic nitrogen source; still further, the inorganic nitrogen source may be urea. The research of the invention finds that yeast extract is not needed to be added when the starch syrup B is used as the raw material for 3-hydroxy butanone fermentation, and the yield and the conversion rate of the 3-hydroxy butanone are obviously higher than those of the fermentation using glucose as the raw material.
In another embodiment of the present invention, the fermentation medium comprises the above starch sugar solution and an inorganic nitrogen source, wherein the inorganic nitrogen source is urea;
in another embodiment of the present invention, the initial concentration of glucose in the fermentation medium is controlled to be 120-160g/L, and the urea concentration is controlled to be 1-2 g/L.
In yet another embodiment of the present invention, the 3-hydroxybutanone producing strain is preferably bacillus, more preferably bacillus subtilis; in the examples of the present invention, the 3-hydroxybutanone producing species was Bacillus subtilis BS 168D.
In another embodiment of the present invention, the process further comprises collecting and purifying 3-hydroxy butanone; specifically, the specific collection and purification method comprises the following steps: collecting 3-hydroxy butanone fermentation liquor, carrying out fractionation treatment, adding salt into the obtained 3-hydroxy butanone fraction, extracting, rectifying, separating and extracting to obtain 3-hydroxy butanone crystals; specific purification methods can also be performed with reference to CN 201310426868.3.
In another embodiment of the present invention, the fractionation treatment method includes: collecting the fraction of 3-hydroxy butanone fermentation liquor by adopting a reduced pressure distillation or multi-effect concentration mode, wherein the distillation temperature is controlled to be 55-75 ℃; collecting the fraction accounting for 75-85% of the total volume of the fermentation liquid. In the invention, 3-hydroxy butanone and water are azeotroped, 3-hydroxy butanone in fermentation liquor can be collected through reduced pressure distillation or multi-effect concentration, and other unfermented components such as pentosan and the like brought by the starch B raw material are enriched in the bottom distillate, so that the extraction cost of the 3-hydroxy butanone is not increased compared with that of a glucose raw material.
In another embodiment of the invention, the 3-hydroxybutanone prepared by the process for producing 3-hydroxybutanone by using wheat B starch is provided.
The invention is further illustrated by the following examples, which are not to be construed as limiting the invention thereto. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
Experimental Material
Experimental strains: bacillus subtilis BS168D (2, 3-butanediol dehydrogenase gene (bdhA) deleted Bacillus subtilis 168 engineered strain) (Junjunjunjun Zhang, Xiaoying Zhao, Jianxiang Zhang, Chen Zhao, Jianjun Liu, Yanjun Tian & Liping Yang (2017) efficiency of deletion of 2, 3-butanol dehydrogenase gene (bdhA) on acetic acid product of Bacillus subtilis, prepared Biochemistry and Biotechnology 47:8, 761-767). The above strains are available to the public from research and design institute of food fermentation industry in Shandong province.
B, obtaining starch: centrifuging fresh wheat starch slurry generated in the production process of wheat gluten by using a centrifuge at the rotating speed of 3000 for 2 seconds, removing supernatant, and collecting the intermediate flowable slurry part for preparing starch sugar solution.
Preparing sugar liquor: adding water into the separated and washed starch for size mixing, adjusting the concentration of the starch size to be 10-15 Be, adding a proper amount of amylase according to the total amount of the starch for sugar preparation by a two-enzyme method (the amount of the enzyme preparation is added according to the amount recommended by a manufacturer), after the saccharification is finished, performing centrifugal separation, and collecting a clear liquid part for 3-hydroxy butanone fermentation.
Seed culture medium: 50g/L of glucose, 10g/L of yeast extract, 10g/L of corn steep liquor, 5g/L of sodium chloride and drinking water, and the pH value is 7.0.
3-hydroxybutanone fermentation
Taking starch sugar solution, adjusting to proper glucose concentration (120-. Control experiments were made with oral glucose. The culture medium is sterilized and inoculated with a strain for fermentation.
And (3) shaking flask fermentation: the liquid loading of a 500ml triangular flask is 50ml, the inoculation amount is 5%, and the culture is carried out for 3-4 days under the optimal culture condition.
Fermentation in a fermentation tank: the fermentation adopts a laboratory scale (5-50L) fermentation tank, and the proper temperature and relative dissolved oxygen concentration are controlled according to requirements in the fermentation process. Culturing until the residual glucose is reduced to below 0.1g/L, and finishing fermentation.
EXAMPLE 1 Effect of the Medium on 3-Hydroxybutanone fermentation
The experiment uses glucose as a control to carry out a 3-hydroxy butanone fermentation experiment, the influence of nitrogen source addition on the fermentation of the 3-hydroxy butanone in the wheat B starch syrup is examined, and the result is shown in the following table:
the experimental result shows that yeast extract and corn steep liquor are not needed to be added when the starch syrup B is used as the raw material for carrying out 3-hydroxy butanone fermentation, and the yield and the conversion rate of the 3-hydroxy butanone are obviously higher than those of glucose.
Example 2 fermentation of 50L Using starch syrup B as a starting Material
Preparing starch sugar from fresh wheat B starch slurry by conventional double-enzyme sugar preparation process, centrifuging after saccharification to remove solid residue, washing the residue with water of the same volume, centrifuging, mixing the centrifugally separated clear liquids, adjusting appropriate glucose concentration, adding into a fermentation tank, and sterilizing. An additional 2g/L urea (separately sterilized, added prior to inoculation) was added. Inoculating according to the inoculation amount of 5%, wherein the initial sugar concentration after inoculation is 138g/L, and controlling the appropriate temperature and relative dissolved oxygen concentration during the fermentation process. After culturing for 58 hours, the glucose concentration was measured to fall below 0.1g/L, and the fermentation was terminated. 35L of fermentation liquor is collected together, and the content of the 3-hydroxy butanone is measured to be 53.2 g/L.
And (3) carrying out reduced pressure distillation on 10L of the fermentation liquor at 70-80 ℃, collecting 8.3L of fraction, measuring the content of 3-hydroxybutanone in the fraction to be 55.6g/L, and determining the recovery rate of 3-hydroxybutanone to be 86.7%.
It should be noted that the above examples are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the examples given, those skilled in the art can modify the technical solution of the present invention as needed or equivalent substitutions without departing from the spirit and scope of the technical solution of the present invention.
Claims (8)
1. A process method for producing 3-hydroxy butanone by using wheat B starch is characterized by comprising the following steps: preparing sugar from wheat B starch to obtain starch sugar liquid, preparing a fermentation culture medium by taking the starch sugar liquid as a raw material, and inoculating a 3-hydroxy butanone production strain into the fermentation culture medium for fermentation to produce 3-hydroxy butanone; the fermentation medium also comprises an inorganic nitrogen source;
wherein the 3-hydroxy butanone production strain is bacillus subtilis, and the inorganic nitrogen source is urea.
2. The process of claim 1, wherein said wheat B-starch is selected from the group consisting of B-starch slurry isolated during vital wheat processing and B-starch products obtained by direct drying of B-starch.
3. The process method of claim 1, wherein the wheat B starch is added with water and is subjected to size mixing to obtain starch slurry, the concentration of the starch slurry is controlled to be 10-15 Be, and then a 'double-enzyme method' sugar preparation process is adopted to prepare a starch sugar solution.
4. The process of claim 1, wherein the initial concentration of glucose in the fermentation medium is controlled to be 120-160g/L and the urea concentration is controlled to be 1-2 g/L.
5. The process of claim 1, wherein the 3-hydroxybutanone producing species is bacillus subtilis BS 168D.
6. The process of claim 1, further comprising collecting and purifying the 3-hydroxybutanone.
7. The process method of claim 6, wherein the specific collection and purification method comprises: collecting 3-hydroxy butanone fermentation liquor, fractionating, adding salt, extracting, rectifying, separating and extracting to obtain 3-hydroxy butanone crystal.
8. The process of claim 7, wherein the specific fractionation treatment method comprises: collecting the fraction of 3-hydroxy butanone fermentation liquor by adopting a reduced pressure distillation or multi-effect concentration mode, wherein the distillation temperature is controlled to be 55-75 ℃; collecting the fraction accounting for 75-85% of the total volume of the fermentation liquid.
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| CN202010413118.2A CN111607622B (en) | 2020-05-15 | 2020-05-15 | Process method for producing 3-hydroxy butanone by using wheat B starch |
| PCT/CN2020/133657 WO2021227453A1 (en) | 2020-05-15 | 2020-12-03 | Processing method for producing 3-hydroxybutanone by means of using wheat b starch |
| NL2028177A NL2028177B1 (en) | 2020-05-15 | 2021-05-10 | Process for producing acetoin by utilizing wheat b-starch |
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| CN115124414A (en) * | 2022-05-20 | 2022-09-30 | 山东省食品发酵工业研究设计院 | Preparation method and application of high-optical-purity 3-hydroxy butanone |
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| CN100455658C (en) * | 2007-01-29 | 2009-01-28 | 山东省食品发酵工业研究设计院 | Bacillus subtilis capable of producing high purity 3-hydroxy butanone |
| CN101049133B (en) * | 2007-04-26 | 2011-07-06 | 华南理工大学 | Method for preparing nitrogen contained syrup in use for ferment through wheat beta starch multienzyme method |
| CN101161818B (en) * | 2007-09-29 | 2010-08-11 | 上海天之冠可再生能源有限公司 | Method for producing acetone-butanol by partly substituting for corn fermentation |
| CN101294143B (en) * | 2008-06-20 | 2010-11-03 | 南京工业大学 | Bacterial strain for preparing 3-hydroxy butanone and uses thereof |
| CN103582787B (en) * | 2011-05-18 | 2016-09-14 | 斯科尔生物燃料公司 | The production technology of the volatile fermentation products of solar energy auxiliary |
| CN102352396B (en) * | 2011-09-01 | 2013-11-13 | 河南飞天农业开发股份有限公司 | Method of producing functional starch sugar through wheat starch |
| CN102899364A (en) * | 2012-09-14 | 2013-01-30 | 日照鲁信金禾生化有限公司 | Method for production of citric acid through fermentation of wheat B-starch |
| CN103333842B (en) * | 2013-07-10 | 2015-03-04 | 山东省食品发酵工业研究设计院 | Bacillus subtilis producing 3-hydroxybutanone and application thereof |
| CN103483167B (en) * | 2013-08-27 | 2016-06-08 | 山东鲁北药业有限公司 | A kind of purification process of 3-Hydroxybutanone |
| CN103524315B (en) * | 2013-09-18 | 2014-12-31 | 山东省食品发酵工业研究设计院 | Method of extracting, distilling and separating 3-hydroxyl butanone from fermentation liquor by adding salts |
| CN104328099A (en) * | 2014-11-13 | 2015-02-04 | 河南天冠纤维乙醇有限公司 | Method for producing beta-glucanase by using recombined pichia pastoris |
| CN105624219A (en) * | 2016-03-07 | 2016-06-01 | 江苏国信协联能源有限公司 | Method for producing citric acid by fermenting wheat starch milk |
| CN105820988B (en) * | 2016-06-08 | 2019-06-11 | 中国石油大学(华东) | One bacillus and its application in hot fermentation production 3-hydroxy-2-butanone and 2,3- butanediol |
| CN107129959B (en) * | 2017-06-28 | 2020-12-18 | 广西科学院 | Construction method and application of genetic engineering strain for producing (R) -acetoin |
| CN110305913B (en) * | 2019-06-28 | 2022-04-12 | 山东省食品发酵工业研究设计院 | Method for producing tetramethylpyrazine from sugar raw material |
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| CN111607622A (en) | 2020-09-01 |
| WO2021227453A1 (en) | 2021-11-18 |
| NL2028177A (en) | 2021-11-23 |
| NL2028177B1 (en) | 2022-03-18 |
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